Retinoid X Receptor Gamma DNA-Binding ELISA Kit (OKAG00458)

Catalog No: OKAG00458

Size:12x8WELLSTRIPS

Retinoid X Receptor γ DNA-Binding ELISA detects active Retinoid X Receptor γ in Jurkat and Hela Nuclear Extract. The Jurkat and Hela cells were grown 3 days respectively in RPMI 1640 and DMEM with 10% FBS and harvested for nuclear extract which is tested on the plate coated with wild type (wt) and mutated oligonucleotides.

Product Info

• Predicted Species Reactivity: Human, Mouse
• Product Format: 12 x 8-Well Microstrips
• Application: Enzyme-linked immunosorbent assay
• ELISA Kit Detection Method: Colorimetric 450 nm
• ELISA Kit Principle: The Aviva DNA-Binding ELISA Kit contains components necessary for detection of active transcription factors in eukaryotic nuclear or cell lysates. This particular immunoassay utilizes the qualitative technique of an indirect ELISA. Streptavidin is bound to the immunoassay plate and specific biotinylated double-stranded (dsDNA) oligonucleotides are then added to bind to the streptavidin via a high affinity biotin-streptavidin interaction. After subsequent blocking of extraneous binding sites in each well, the sample containing the target of interest can be added. Primary antibody is added to bind activated transcription factors bound to the dsDNA oligonucleotide, which has been immobilized via the plate-coated streptavidin. A HRP-conjugated secondary antibody specific for rabbit IgGs is added, which allows for specific binding to the Primary Antibody, and consequently colorimetric detection upon addition of the TMB substrate.
  For color development, TMB (3, 3’, 5, 5’-Tetramethylbenzidine) is added to each well. After addition of the substrate, a peroxidase catalyzed reaction will produce a blue TMB Diimine product that is proportional to the target concentration in the sample. Color development is quenched by addition of Stop Solution, or 2 N Sulfuric Acid, which turns the solution yellow. The absorbance can then be read by a spectrophotometer at 450 nm and subsequently allowing for determination of the target concentration in the sample.
  
• ELISA Kit Component:
  • dsDNA Oligonucleotide Coated Microplate: 96 Wells (12 x 8 Strips)Microstrips
  • 100X Primary Antibody: 100 uL
  • 100X Primary Phospho-Antibody: 100 uL
  • HRP-Conjugated Anti-Rabbit IgG Antibody: 2 x 6 mL
  • Nuclear Lysate Positive Control: Lyophilized
  • Wild-Type Consensus dsDNA Oligonucleotide: 20 uL
  • Mutant Consensus dsDNA Oligonucleotide: 20 uL
  • 10X Wash Buffer: 50 mL
  • 2X Binding Buffer: 12 mL
  • Primary Antibody Diluent: 12 mL
  • 100X Protease and Phosphatase Inhibitors: 100 uL
  • Nuclear Wash Buffer: 12 mL
  • Cytoplasmic Extraction Buffer: 6 mL
  • Nuclear Extraction Buffer: 6 mL
  • Ready-to-Use Substrate: 12 mL
  • Stop Solution: 12 mL
  
  OMIM: 180247
  UniGene: Hs.26550
  Note: The product is for research use only, not for use in diagnostic or therapeutic procedures.
• Reconstitution and Storage: Store as indicated in product manual.
• Specificity: The Retinoid X Receptor γ DNA-Binding ELISA Kit detects endogenous levels of Retinoid X Receptor γ protein.
• Assay Info: Assay Type: Qualitative DNA Binding ELISA

Target Info

• Gene Symbol: RXRG
• Gene Full Name: retinoid X receptor gamma
• Alias Symbols: RXRC, NR2B3
• NCBI Gene Id: 6258
• Protein Name: retinoic acid receptor RXR-gamma
• Description of Target: This gene encodes a member of the retinoid X receptor (RXR) family of nuclear receptors which are involved in mediating the antiproliferative effects of retinoic acid (RA). This receptor forms dimers with the retinoic acid, thyroid hormone, and vitamin D receptors, increasing both DNA binding and transcriptional function on their respective response elements. This gene is expressed at significantly lower levels in non-small cell lung cancer cells. Alternatively spliced transcript variants have been described.
• Uniprot ID: P48443